Retrovirus-mediated expression of preprosomatostatin in rat pituitary GH3 cells

Targeting of somatostatin to the regulated secretory pathway

N. Fleischer, H. Liker, T. Stoller, R. D'Ambra, D. Shields

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Somatostatin (SRIF) is a 14-amino acid peptide hormone that is synthesized as part of a larger precursor, prepro-SRIF, consisting of a signal peptide and a proregion of 80-90 amino acids; mature SRIF is located at the carboxyl-terminus of the precursor. We have used a recombinant retroviral expression vector encoding anglerfish prepro-SRIF-I to infect rat pituitary GH3 cells. The aim of these studies was to investigate the intracellular storage and secretion of the total pool of endogenous GH compared to that of SRIF. Several clonal lines of GH3 cells expressing high or low levels of SRIF were treated with THR, forskolin, or depolarizing concentrations of potassium, and the levels of intracellular and secreted GH or SRIF were determined using highly sensitive RIAs. Approximately 65% of the total GH was secreted basally, whereas less than 20% of the SRIF-immunoreactive material was basally secreted. Forskolin treatment or potassium depolarization stimulated GH release, but only about 50% above basal levels. In contrast, SRIF secretion was stimulated approximately 5-fold in response to these secretagogues. Based on its lower basal rate of secretion compared to GH and its enhanced release in response to a variety of secretagogues, we conclude that the heterologously expressed SRIF is preferentially targeted to the regulated pathway in GH3 cells.

Original languageEnglish (US)
Pages (from-to)1652-1658
Number of pages7
JournalMolecular Endocrinology
Volume3
Issue number10
StatePublished - 1989

Fingerprint

Somatostatin-Secreting Cells
Secretory Pathway
Retroviridae
Colforsin
Potassium
Amino Acids
Peptide Hormones
Protein Sorting Signals
Somatostatin

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Retrovirus-mediated expression of preprosomatostatin in rat pituitary GH3 cells : Targeting of somatostatin to the regulated secretory pathway. / Fleischer, N.; Liker, H.; Stoller, T.; D'Ambra, R.; Shields, D.

In: Molecular Endocrinology, Vol. 3, No. 10, 1989, p. 1652-1658.

Research output: Contribution to journalArticle

Fleischer, N. ; Liker, H. ; Stoller, T. ; D'Ambra, R. ; Shields, D. / Retrovirus-mediated expression of preprosomatostatin in rat pituitary GH3 cells : Targeting of somatostatin to the regulated secretory pathway. In: Molecular Endocrinology. 1989 ; Vol. 3, No. 10. pp. 1652-1658.
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